Rapid reconstitution for lyophilized-pharmaceutical suspensions

ABSTRACT

A method of preparing and reconstituting a sterile, lyophilized pharmaceutical active for rapid reconstitution by evacuating a lyophilized pharmaceutical active-containing container until the pressure within the container is less than about 300 Torr and hermetically sealing the evacuated container. The sterile, lyophilized pharmaceutical active can be prepared by flash freezing a pharmaceutical active-containing composition then lyophilizing the composition. The hermetically sealed lyophilized pharmaceutical active can be reconstituted by adding at least the total volume of liquid necessary for reconstitution of the sterile, lyophilized pharmaceutical active to the sterile, lyophilized pharmaceutical active, sealed under a pressure of less than about 300 Torr, in less than about 10 seconds to yield, within about 5 minutes, an administrable pharmaceutical active-containing composition. One aspect of the herein described sterile, lyophilized pharmaceutical active is a packaged sterile pharmaceutical active comprising an evacuated, hermetically sealed container having disposed therein a sterile, lyophilized pharmaceutical active, sealed under a pressure of less than about 300 Torr.

CROSS-REFERENCE TO RELATED APPLICATION

The benefit of U.S. Provisional Patent Application 61/290,998, filed Dec. 30, 2009, is hereby claimed, and the disclosure thereof is incorporated herein by reference in its entirety.

FIELD OF THE DISCLOSURE

The invention relates to the pharmaceutical and medical field and provides a packaged lyophilized pharmaceutical active and a method of producing the packaged lyophilized pharmaceutical active. Specifically, the invention provides for the rapid reconstitution of a lyophilized pharmaceutical active with minimal foaming.

BACKGROUND

Lyophilization is used to prepare pharmaceutical actives with higher stability, broader temperature tolerance, and longer shelf-life than comparable aqueous solutions. Typically, pharmaceutical actives are dried to water contents of less than 5 wt. % through sublimation and desorption.

The reconstitution of lyophilized pharmaceutical actives that contain proteins or polypeptides by the addition of water, leads to the formation of a foam. The foam is thought tooccur from the interaction of the amphiphilic protein or polypeptide with the reconstituting solvent, typically water. The foaming of the lyophilized pharmaceutical active poses problems for medical practitioners and researchers. For example, in double blind studies placebos of lyophilized pharmaceuticals need to appear identical to the active therefore a placebo must foam identically to the active. See e.g., U.S. Pat. No. 6,242,423. The reduction or elimination of foaming would benefit both the medical practitioner by saving time and the researcher by reducing the complication of formulating a placebo for a double blind study.

SUMMARY OF THE INVENTION

Described herein, in the preferred embodiment, is a lyophilized pharmaceutical active stored under a pressure of less than 300 Torr (40 kPa) that can be rapidly reconstituted for in vivo use.

One aspect of the materials and methods described herein is a process for making a sterile, lyophilized pharmaceutical active. This process includes evacuating a container that contains the sterile, lyophilized pharmaceutical active and hermetically sealing the container.

Another aspect of the materials and methods described herein is a process for preventing Ostwald ripening growth during the preparation of a sterile, lyophilized pharmaceutical active. This process includes flash freezing a sterile pharmaceutical active containing composition, lyophilizing the composition, and sealing the sterile, lyophilized pharmaceutical active under a pressure of less than about 300 Torr (40 kPa).

Yet another aspect of the materials and methods described herein is a process for reconstituting a sterile, lyophilized pharmaceutical active. This process includes adding a sterile liquid to the sterile, lyophilized pharmaceutical active to yield an administrable pharmaceutical active-containing composition within about 5 minutes.

Still another aspect of the materials and methods described herein is a packaged sterile pharmaceutical active. This package includes a sterile, lyophilized pharmaceutical active hermetically sealed within a container.

DETAILED DESCRIPTION

Herein, ranges may be expressed as from “about” or “approximately” one particular value and/or to “about” or “approximately” another particular value. When such a range is expressed, another embodiment includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. Additionally, compilations of parts of or areas in devices are at times designated specific regions, these regions are described based on the theorized primary event occurring in the designated region. Regions can and often overlap and other events can and likely occur within the specifically designated regions.

The article and methods described herein generally relates to the preparation, storage and reconstitution of pharmaceutical actives. An aspect of the reconstitution of pharmaceutical actives is the amount of time necessary for the reconstitution, with multi-step and/or prolonged reconstitution processes being burdensome on an administrating medical practitioner. Embodiments of the herein described methods and article can be selected to reduce the medical practitioner's time commitment to the reconstitution process and thereby the medical practitioner can more readily care for the patient.

One aspect of the herein described article and methods is the preparation of a sterile, lyophilized pharmaceutical active (“LPA”) for rapid reconstitution, comprising evacuating a lyophilized pharmaceutical active-containing container until the pressure within the container is less than about 25 Torr (3333 Pa); and hermetically sealing the evacuated container. Another aspect of the herein described methods is the prevention of Ostwald ripening growth by flash freezing during the preparation of the sterile, LPA. Still another aspect of the herein described methods is the rapid reconstitution of the sterile, LPA. Yet another aspect of the herein described article is the packaged sterile, LPA, for example, prepared by any one of the methods described herein.

Generally, the preparation of the sterile, LPA described herein involves the lyophilization and storage of a pharmaceutical active-containing composition, often containing a solvent or liquid, e.g., water. In addition, the pharmaceutical active can be in the formof a solution, a suspension, or an emulsion. The pharmaceutical active can be lyophilized by placing a pharmaceutical active-containing composition in a freeze dryer or a lyophilizer, and lyophilizing. One procedure for lyophilizing a pharmaceutical active-containing composition includes continuously pulling a vacuum (e.g. to less than about 1 Torr (133 Pa), preferably about 0.01 to about 1 Torr, more preferably about 60 to about 300 mTorr (8 to 40 Pa), even more preferably to about 100 mTorr (13 Pa) or less) on a pharmaceutical active-containing composition that was precooled and maintained at a temperature of less than about −20° C., more preferably a temperature of about −20° C. to about −70° C., for example a temperature of about −40° C.

A lyophilization of a pharmaceutical active-containing composition can be divided into a plurality of drying cycles, e.g., the drying cycles can include a primary drying cycle and a secondary drying cycle. The primary drying cycle can be a period of time that depends on the removal of an amount of frozen solvent, e.g., the majority, from the pharmaceutical active-containing composition. The plurality of drying cycles can include a primary to secondary drying cycle transition which can include raising the temperature of the pharmaceutical active-containing composition, e.g., at about 0.5° C. per minute to about −20° C., while continuously pulling a vacuum. The secondary drying cycle can be a period of time that depends on the removal of an amount of residual solvent in the pharmaceutical active-containing composition, e.g., that not removed in the primary drying cycle, if any, and which would be apparent to one of ordinary skill. The plurality of drying cycles can further include a secondary to tertiary drying cycle transition that can include raising the temperature of the pharmaceutical active-containing composition, at about 0.5° C. per minute to about 30° C., while continuously pulling a vacuum. A tertiary drying cycle can be a period of time, that is apparent to one of ordinary skill, that includes continuously pulling vacuum at a temperature above the melting point of a solvent that was included in the pharmaceutical active-containing composition prior to lyophilization.

An added benefit can be obtained if Ostwald ripening growth is prevented during the preparation of the sterile, LPA. Preferably, the pharmaceutical active-containing composition is flash frozen prior to lyophilization. The pharmaceutical active-containing composition can be flash frozen by, for example, submerging a container holding the composition in a cryogenic liquid, e.g., liquid nitrogen. Alternative methods exist for the flash freezing of the composition and will be available to one of ordinary skill. Preferably, the composition is frozen at a sufficient rate (e.g. within about 60 seconds) to prevent Ostwald ripening growth. When the pharmaceutical active-containing composition is cryogenically flash frozen the composition can be subsequently warmed within the lyophilization chamber (e.g. to a temperature of about −40° C.) while exposed to a vacuum (e.g. to less than about 1 Torr (133 Pa), preferably about 0.01 to about 1 Torr, more preferably about 60 to about 300 mTorr (8 to 40 Pa), even more preferably to about 100 mTorr (13 Pa) or less).

Generally, the storage of the LPA involves disposing the LPA in a sealable container and preferably sealing the container. Preferably, after lyophilization, the LPA is disposed within a hermetically sealable container; the container is evacuated, preferably, to a pressure less than about 300 Torr, more preferably less than about 200 Torr, even more preferably less than about 100 Torr, still more preferably less than about 50 Torr, even still more preferably less than about 25 Torr (3333 Pa), yet more preferably less than about 10 Torr (1333 Pa), and yet still more preferably less than about 5 Torr (667 Pa) or 4 Torr (533 Pa); and the container is then hermetically sealed. One of ordinary skill will comprehend methods for storing a material within a container. These methods include and are not limited by the methods of 1) storing the LPA in a sealable container, evacuating the container, and then hermetically sealing the container; 2) sealing the LPA in a container, evacuating the container, and then applying a hermetic seal; or 3) evacuating a chamber containing the LPA and the container, and then hermetically sealing the LPA inside the container. Optionally, the sealed container contains a dissolution aid, for example a dissolution aid can be a impeller, a magnetic stirrer, and/or other inert solids like glass beads, polyethylene beads, and ceramic beads.

The sealable container can be a rigid container, a non-rigid container, or a variable-volume container. Examples of rigid containers include ampoules, vials, tubes, bottles, and the like; for example made of glass, plastic, and the like. Non-rigid containers include hermetically sealable plastic bags. When the sealable container is a non-rigid container one of ordinary skill would recognize that the pressure within the container after equilibration is typically not the same pressure as when the container was evacuated, e.g., the container deforms to equilibrate the external pressure and the internal pressure; a non-rigid container sealed under a pressure of less than about 25 Torr (3333 Pa) will equilibrate to a higher internal pressure, approaching or achieving the external pressure. Variable-volume containers include for example syringes. While syringes hermetically sealed under vacuum may decrease the internal volume within the syringe, syringes with barrel-locking mechanisms may be used to maintain a minimum volume within the syringe and thereby a known internal pressure. One of ordinary skill in the art will comprehend how to hermetically seal the sealable container. Rigid containers are preferred.

The methods described above will yield a packaged sterile, LPA. Preferably, the packaged sterile, LPA comprises a LPA disposed within a hermetically sealable container. More preferably, the LPA is sealed within the container at a pressure of less than about 300 Torr, more preferably less than about 200 Torr, even more preferably less than about 100 Torr, still more preferably less than about 50 Torr, even still more preferably less than about 25 Torr (3333 Pa), yet more preferably less than about 10 Torr (1333 Pa), and yet still more preferably less than about 5 Torr (667 Pa) or 4 Torr (533 Pa). One of ordinary skill in the art will recognize that the pressure in the hermetically sealed container is dependent on the rigidity of the container, therefore the pressure within the container may be different from the pressure at which the container was sealed.

The reconstitution of the pharmaceutical active by the addition of a solvent for the pharmaceutical active is preferably carried out under reduced pressure. One of ordinary skill in the art will recognize that there are multiple methods for the addition of a solvent to a material stored under a reduced pressure. Herein, all methods for the addition of the solvent to the pharmaceutical active are applicable so long as the addition occurs with minimum addition of a gas. These methods include the cannulae transfer of the solvent, the syringe transfer of the solvent, addition of the solvent in a vacuum chamber, vacuum distillation of the solvent into the pharmaceutical active containing container, and the like. More preferably, the addition of the solvent occurs without the formation of bubbles or foam in the pharmaceutical active-containing container. Still more preferably, the solvent is degassed prior to the addition to the pharmaceutical active.

The reconstitution of the pharmaceutical active is preferably rapid. Herein, the reconstitution of known pharmaceutical actives preferably occurs in 50% less time than the time necessary under the current and accepted procedures for the reconstitution of the same pharmaceutical actives. More preferably, the reconstitution of the pharmaceutical active occurs as a single step through the addition of the total volume of solvent necessary for reconstitution of the pharmaceutical active. Preferably, the total volume of solvent is added in less than about 60 seconds, more preferably less than about 30 seconds, even more preferably less than about 20 seconds, and still more preferably less than about 10 seconds; thereafter the reconstituted pharmaceutical active-containing composition is preferably administrable within about 10 minutes, more preferably within about 5 minutes, and even more preferably within about 2.5 minutes.

The pharmaceutical active is one that can be lyophilized and reconstituted. In one embodiment, the pharmaceutical active includes an albumin. Typically, lyophilized albumin compositions foam upon reconstitution with water due to the interaction of the amphiphilic albumin and the water. The foaming of lyophilized albumin forces prolonged reconstitution procedures to prevent the injection or biological application of the foamed albumin, thereby preventing rapid reconstitution and administration. The foaming of the reconstituted albumin is especially detrimental to the rapid reconstitution and administration of pharmaceutical actives intended for human use. When intended for human use the albumin is preferably a human serum albumin. More preferably the pharmaceutical active has nanoparticles of human serum albumin. The amount of albumin in the pharmaceutical active can vary. Preferably, the pharmaceutical active has at least about 50 wt. % of the albumin, more preferably about 75 wt. %, and even more preferably about 85 wt. %. When the pharmaceutical active comprises an albumin the active can further comprise a drug or drugs that bind to an albumin. Examples of drugs that bind to human serum albumin are known to one of ordinary skill and include ontological, imaging, anti-inflammatory, and antiallergic drugs. A preferable ontological drug that binds to human serum albumin is paclitaxel.

EXAMPLES

Several lyophilization and reconstitution studies were performed to test the invention using: 1) ABRAXANE vials, 2) homogenized 0.3% paclitaxel/albumin formulation, 3) 5% BSA (bovine serum albumin), and 4) 3% BSA. ABRAXANE is a nanoparticle suspension containing approximately 4.5% HSA (human serum albumin) and 0.5% paclitaxel after reconstituted with 20 mL of the diluent. Thus, 90% of the content of the ABRAXANE vials is HSA which is a protein. As such, HSA contributes the major physical characteristics, such as texture of the lyophilized vial, reconstitution rate, and foaming, of the ABRAXANE product. Thus placebo vials, i.e. vials containing HSA without paclitaxel, will give a close representation of the above physical characteristics. For convenience, BSA was used as the placebo for the testing, since pure BSA in solid form is more available than HSA. The 5% BSA was used to mimic the ABRAXANE with 20 mL of fill volume for lyophilization, and 3% BSA was used as the placebo to mimic 33 mL fill volume for lyophilization. The studies were performed per the following experimental designs:

-   -   1) Reconstituted the original ABRAXANE vial with 20 mL pure         water and recorded the reconstitution time following the         procedure described in the package insert of the product.     -   2) Pulled a vacuum on the ABRAXANE vial, to approximately 4 Torr         (533 Pa) using a LYOSTAR II dryer, followed by reconstitution         with 33 mL water to study the effect of vacuum on the         reconstitution rate.     -   3) Divided the above reconstituted vial into 4 vials, 4 mL each         into 10 mL vials, followed by lyophilization and sealing under a         high vacuum of 4 Torr (533 Pa) to 5 Torr (667 Pa). Studied the         reconstitution time of the resulting vials. The purpose of         dividing the content in the ABRAXANE vial was due to the high         cost of ABRAXANE. The results from these small vial studies gave         a close indication of the physical characteristics of the actual         size of the ABRAXANE vial.     -   4) Prepared 5% BSA and filled 4 mL each into 10 mL vials,         followed by lyophilization and reconstitution, and compared the         results with those in Step 2.     -   5) Lyophilized approximately 30 mL of 0.3% paclitaxel/albumin         nanoparticle suspension in 50 mL vials, followed by         lyophilization and reconstitution.     -   6) Prepared 3% BSA and filled 33 mL each into 50 mL vials,         followed by lyophilization and reconstitution, and compares the         results with those in Step 4.

The results of the above studies are presented below.

Example 1 Lyophilization and Reconstitution Studies for 5% BSA (Bovine Serum Albumin), 5 mL Fill into 10 mL Vials

The purpose of using 4 mL fill of ABRAXANE in 10 mL vials was due to the high cost of ABRAXANE. The small vial studies generated more replicates for data analysis.

The lyophilization cycle parameters are:

-   -   1) Cool the shelf to −40° C. and hold for 80 minutes.     -   2) Pull vacuum to 100 mTorr (133 Pa).     -   3) Hold at −40° C., 100 mTorr (133 Pa) for 30 minutes.     -   4) Ramp 0.5° C. to −20° C., 100 mTorr (133 Pa).     -   5) Primary drying: Hold at −20° C., 100 mTorr (133 Pa) for         approximately 2300 minutes.     -   6) Ramp 1° C./min to 30° C., 100 mTorr (133 Pa).     -   7) Secondary drying: Hold at 30° C., 100 mTorr (133 Pa) for         approximately 330 minutes.

After lyophilization, the vials were sealed at approximately 5 Torr (667 Pa). The reconstitution was performed by pointing the needle at the cake to allow water to quickly absorbed by the cake.

It was observed minimal amount of foam produced, probably due to the fact that the vials were sealed at a high vacuum of approximately 5 Torr (667 Pa). The vial was gently rotated by fingers, and it was observed complete dissolution of the cake approximately 4 minutes from the time when water was injected into the vial.

Comparative Example 1 Reconstitution of the Original ABRAXANE Vial with 20 mL Water

The purpose of the study was to determine the reconstitution time and foaming phenomenon of the ABRAXANE vial which contained very little vacuum.

Reconstitution time Comment Approximately 20 min The reconstitution procedure provided in the package insert was carefully followed to avoid foaming.

As seen, approximately 20 minutes was required to reconstitute the vial with a very careful control of the reconstitution procedure described in the package insert.

Example 2 Lyophilization and Reconstitution Studies for Reconstituted ABRAXANE Vial (4 mL Filled into 10 mL vials) and 5% BSA (5 mL Fill into 10 mL Vials)

The purpose of the study was to investigate the lyophilization and reconstitution characteristics of the reconstituted ABRAXANE vials and the placebo vials of 5% BSA.

The lyophilization cycle parameters are:

-   -   1) Cool the shelf to −40° C. and hold for 90 minutes.     -   2) Pull vacuum to 100 mTorr (133 Pa).     -   3) Hold at −40° C., 100 mTorr (133 Pa) for 20 minutes.     -   4) Ramp at 0.5° C./min to −20° C., 100 mTorr (133 Pa).     -   5) Hold at −20° C., 100 mTorr (133 Pa) for approximately 2400         minutes.     -   6) Ramp at 0.5° C./min to 30° C., 100 mTorr (133 Pa).     -   7) Hold at 30° C., 100 mTorr (133 Pa) for approximately 300         minutes.

After completion of the cycle run, the vials were sealed at approximately 4 Torr 533 Pa) and crimped. The results of reconstitution are discussed below.

(1) Reconstitution Time of 4 mL of Reconstituted ABRAXANE Vials

Sample number Reconstitution time Comments 1 3 min 35.41 sec foaming was minimal 2 3 min 36.05 sec foaming was minimal 3 3 min 34.72 sec foaming was minimal 4 2 min 41.89 sec foaming was minimal 5 3 min 25.21 sec foaming was minimal

(2) Reconstitution Time for 5% BSA

Sample number Reconstitution time Comments  6  4 min 25.87 sec foaming was minimal  7 3 min 2.87 sec foaming was minimal  8  0 min 33.91 sec foaming was minimal  9 4 min 6.22 sec foaming was minimal 10  1 min 44.75 sec foaming was minimal 11  3 min 46.03 sec foaming was minimal 12  2 min 50.19 sec foaming was minimal

As seen, the reconstitution time for all vials are very short.

(3) 5% BSA with Glass Beads

Sample number Reconstitution time Comments 13 1 min 39.75 sec foaming was minimal 14 1 min 55.41 sec foaming was minimal 15 0 min 33.72 sec foaming was minimal 16 1 min 55.41 sec foaming was minimal 17 1 min 36.34 sec foaming was minimal

The above reconstitution times are even shorter than those of 5% BSA without glass beads in (2).

Example 3 Quick Freezing, Lyophilization and Reconstitution Studies for Reconstituted ABRAXANE Vial (4 mL filled into 10 mL Vials) and 5% BSA (5 mL Filled into 10 mL Vials)

The purpose of quick freezing was to immobilize nanoparticles from aggregation or crystallization due to the Ostwald ripening effect.

Quick Freezing Procedure:

-   -   1) Placed the test-tube rack on a metal tray and placed apiece         of aluminum foil on the bottom of the test tube rack and lifted         the four sides of the foil to cover the bottom half of the rack         for holding the liquid nitrogen.     -   2) Placed the reconstituted vials in the slots of the test tube         rack.     -   4) Placed stoppers on the vials.     -   5) Poured liquid nitrogen into the test tube rack to quick         freeze the vials.     -   6) Started the LyoStar II freeze dryer to precool the shelf to         −40° C.

7) Loaded the frozen vials onto the shelf, and started the following cycle.

Lyophilization Cycle Parameters:

-   -   1) Cool the shelf to −40° C. and hold for 90 minutes.     -   2) Pull vacuum to 100 mTorr (133 Pa).     -   3) Hold at −40° C., 100 mTorr (133 Pa) for 30 minutes.     -   4) Ramp at 0.5° C./min to −10° C., 100 mTorr (133 Pa).     -   5) Primary drying: Hold at −10° C., 100 mTorr (133 Pa) for         approximately 1050 minutes.     -   6) Ramp at 0.5° C./min to 30° C., 100 mTorr (133 Pa).     -   7) Secondary drying: Hold at 30° C., 100 mTorr (133 Pa) for         approximately 530 minutes.

After completion of the cycle, the vials were sealed at approximately 4 Torr (533 Pa) and crimped.

The Results of Reconstitution Studies are Presented Below

(1) ABRAXANE Vials (Injected with 4 mL Pure Water)

Sample number Reconstitution time Comment 18 2 min 3.50 sec foaming was minimal 19  2 min 15.66 sec foaming was minimal 20  2 min 45.25 sec foaming was minimal

(2) 5% BSA Vials Without Glass Beads (Injected with 5 mL Pure Water)

Sample number Reconstitution time Comments 21 59.94 sec foaming was minimal 22 2 min 49.84 sec foaming was minimal 23 4 min 6.45 sec foaming was minimal 24 1 min 47.66 sec foaming was minimal 25 2 min 31.72 sec foaming was minimal 26 4 min 12.45 sec floating cake taking longer to dissolve 27 3 min 35.35 sec foaming was minimal

(3) 5% BSA with Glass Beads (Injected with 5 mL Pure Water)

Sample number Reconstitution time Comments 28 51.84 sec foaming was minimal 29 57.22 sec foaming was minimal 30 2 min 57.03 sec foaming was minimal 31 3 min 49.00 sec foaming was minimal

The above results indicate that the resulting cakes after quick freezing followed by lyophilization also reconstituted rapidly. Thus the combination of quick freezing and vacuum seal can prevent aggregation (or crystallization) of nanoparticles and give a quick reconstitution of the resulting cakes.

Comparative Example 3 Reconstitution of Vials Sealed Under Ambient Pressure

The purpose of this study was to investigate the reconstitution time for samples prepared by the quick freezing and lyophilization procedure described above wherein the samples were sealed under ambient pressure, approximately 760 Torr (101 kPa).

(1) ABRAXANE Vials (Injected with 4 mL Pure Water)

Sample number Reconstitution time Comment 32 3 min 41.66 sec large amount of foaming

(2) 5% BSA Vials without Glass Beads (Injected with 5 mL Pure Water)

Sample number Reconstitution time Comments 33 9 min 30.28 sec large amount of foaming

(3) 5% BSA with Glass Beads (Injected with 5 mL Pure Water

Sample number Reconstitution time Comments 34 4 min 52.93 sec large amount of foaming

The above-described results indicate that the resulting cakes, after quick freezing followed by lyophilization, cannot be reconstituted rapidly if the samples are not sealed under vacuum. As seen, the time necessary for reconstitution ranged from about 60% longer to about 260% longer when the vials were sealed without vacuum compared to sealing under a pressure of approximately 4 Torr (533 Pa).

Example 4 Vacuum Pulled on the Original ABRAXANE Vial, Followed by Reconstitution with 33 mL Water

The purpose of the study was to investigate the effect of vacuum on the reconstitution rate of the original ABRAXANE product vial. The ABRAXANE vial was vacuum pulled to approximately 4 Torr (533 Pa) using LyoStar II dryer and resealed. The purpose of vacuum pull was to minimize foaming during reconstitution. The ABRAXANE vial was then reconstituted with 33 mL of pure water.

Vial number Reconstitution time 35 Approximately 12 minutes and 1.9 sec.

The reconstitution was performed by quickly injecting water through the stopper and pointing at the lumps. It was observed that the reconstitution was shorter than the 20 minutes in Comparative Example 1.

Example 5 Lyophilization and Reconstitution Studies for 0.3% Paclitaxel/Albumin and 3% BSA

The 0.3% paclitaxel/albumin nanoparticle suspension was prepared at Baxter Round Lake facility. The vials contained approximately 30 mL each in 50 mL vials. The placebo of 3% BSA was prepared at Baxter Bloomington facility. The fill volume of the placebo was 33 mL each in 50 mL vials. The vials were lyophilized together using the following cycle parameters:

-   -   (1) Precool the shelf to 5° C.     -   (2) Load the vials to the shelf and install thermocouples.     -   (3) Cool the shelf to −40° C. and hold for approximately 270         min.     -   (4) Start vacuum pump at 100 mTorr (133 Pa) and hold at −40° C.         for 30 min.     -   (5) Primary drying: ramp to −10° C. at 0.5° C./min, at 100 mTorr         (133 Pa), and hold for 135 hours and 37 minutes.     -   (6) Secondary drying: ramp to 30° C. at 1° C./min, and hold for         10 hours.

After completion of the cycle, the vials were sealed at approximately 4 Torr (533 Pa). The following procedure was used to reconstitute the vials.

-   -   (1) Removed the aluminum cap of the vial.     -   (2) Load the syringe with 20 mL o ater, and ensure all air is         removed by priming.     -   (3) Stick the needle through the stopper and point at the cake     -   (4) The water in the syringe will be automatically and quickly         pulled into the vial due to the high vacuum of approximately 4         Torr (533 Pa).     -   (5) As soon as all water is delivered, pull out the needle         quickly to avoid injecting air.     -   (6) Gently swirl or invert the vial to enhance dissolution of         the cake.

As seen, the reconstitution procedure for the vacuum sealed vials is very different than the ABRAXANE product vials, because of foaming can be minimized by the high vacuum.

The following sample IDs are for 3% BSA to mimic the ABRAXANE vials, since the solution in the vials became clear after reconstitution and easier to observe the reconstitution and foam dissipation. The reconstitution times for 3% BSA, 33 mL in 50 mL vials, are presented below:

Sample ID Reconstitution time Comment 36  7 min 23.28 sec foaming was minimal 37  6 min 36.72 sec foaming was minimal 38 10 min 13.38 sec foaming was minimal 39  8 min 58.96 sec foaming was minimal 40  4 min 07.53 sec foaming was minimal Crushed cake by shaking 41  8 min 26.18 sec foaming was minimal

It was found that the crushed cake in sample 40 reconstituted quicker. The cake was crushed by hand, by end-to-end shaking of the vial for a few minutes. The reconstitution times of the intact cake, samples 36, 37, 38, 39, and 41, are still much shorter than that of the ABRAXANE vial. This result further confirmed the effectiveness of vacuum seal on enhancing the reconstitution and minimizing foaming. 

1. A method of preparing a sterile, lyophilized pharmaceutical active for rapid reconstitution, comprising: evacuating a sterile, lyophilized pharmaceutical active-containing container until the pressure within the container is less than about 300 Torr (40 kPa); and hermetically sealing the evacuated container.
 2. The method of claim 1 further comprising: flash freezing a sterile pharmaceutical active-containing composition; lyophilizing the flash frozen sterile pharmaceutical active-containing composition to yield the sterile, lyophilized pharmaceutical active; and thereby preventing Ostwald ripening growth.
 3. The method of claim 2, wherein the lyophilizing of the flash frozen pharmaceutical active-containing composition comprises: storing a flash frozen pharmaceutical active-containing composition in a lyophilization chamber; warming the flash frozen pharmaceutical active-containing composition disposed within the lyophilization chamber to a temperature of less than about −20° C.; pulling a vacuum of less than about 1 Torr (133 Pa) on the lyophilization chamber; warming the flash frozen pharmaceutical active-containing composition disposed within the lyophilization chamber to about −20° C.; and maintaining the vacuum of less than about 1 Torr (133 Pa) on the lyophilization chamber for a period of time constituting a primary drying cycle that corresponds with a removal of a majority of a frozen solvent.
 4. The method of claim 3 further comprising: after the end of the primary drying cycle, then warming the pharmaceutical active-containing composition to about 30° C. and maintaining the vacuum of less than about Ton (133 Pa) for a period of time constituting a secondary drying cycle.
 5. A method of reconstituting a sterile, lyophilized pharmaceutical active comprising: adding a volume of a sterile liquid to a sterile, lyophilized pharmaceutical active, sealed within an evacuated container wherein the pressure within the evacuated container is less than about 300 Torr (40 kPa), in less than about 10 seconds to yield, within about 5 minutes, an administrable pharmaceutical active-containing composition; wherein the volume of liquid added is at least the total volume of liquid necessary for reconstitution of the sterile, lyophilized pharmaceutical active.
 6. The method of claim 5, wherein the sterile, lyophilized pharmaceutical active is one having been lyophilized from a first volume of a liquid containing the pharmaceutical active, and wherein the volume of liquid added for reconstitution is equivalent to the first volume.
 7. The method of claim 5, wherein the liquid is a solvent for the pharmaceutical active.
 8. A packaged sterile pharmaceutical active comprising a sterile, lyophilized pharmaceutical active hermetically sealed within an evacuated container; wherein the pressure within the evacuated container is less than about 300 Torr (40 kPa).
 9. The package of claim 8, wherein the container is selected from the group consisting of a rigid container, a non-rigid container, and a variable-volume container.
 10. The package of claim 8, wherein the sterile, lyophilized pharmaceutical active comprises paclitaxel.
 11. The package of claim 8, wherein the sterile, lyophilized pharmaceutical active comprises an albumin.
 12. The package of claim 11, wherein the pharmaceutical active comprises at least about 50 wt. % of the albumin and wherein the albumin is a human serum albumin.
 13. The package of claim 12, wherein the pharmaceutical active comprises at least about 75 wt. % of the human serum albumin.
 14. The package of claim 13, wherein the pharmaceutical active comprises at least about 85 wt. % of the human serum albumin.
 15. The package of claim 8, wherein the sterile pharmaceutical active-containing composition comprises a nanoparticle of human serum albumin.
 16. The package of claim 8, wherein the pressure within the evacuated container is less than about 100 Torr (13 kPa).
 17. The package of claim 16, wherein the pressure is less than about 25 Torr (3325 Pa).
 18. The package of claim 17, wherein the pressure is less than about 5 Torr (667 Pa). 